Identification of animal and plant species in Foodstuffs using Target GBS assay

FORLANI L.; POSIK D.M.; BRUNO M.C.; OLIVERA L.H.; ZAPPA M.E.; CASTILLO N.S.; BARBISAN G.; VILLEGAS CASTAGNASSO E E; CRESPI J.A.; PERAL GARCIA P.; FERNANDEZ M.E.; GIOVAMBATTISTA G.

Ciudad del Cabo

Congreso; The 39th International Society for Animal Genetics Conference; 2023

ISAG

DNA metabarcoding assay is increasingly used for species authentication in industrialized food. Targeted Genotyping-by-Sequencing (GBS) based on nextgeneration sequencing (NGS) technology allows the detection of multiple species in complex foodstuff matrices in a single run. This study aimed to evaluate the species of origin used in 32 commercially processed foods, replicated thrice, such as broth, salad dressing, sauce, milk, milk powder, hamburger, canned tuna, pepper, etc. In addition, positive (known DNA mixed) and negative controls were added. Ten ng of DNA formeach samples were analyzed using a previously developed AgriSeqTM targeted GBS assay that include 319 multiplexed targets belonging to two mitochondrial (COI and CYTB) and two chloroplast (RBCL and MATK) genes from 177 plant and animal species used in the food industry in Argentina. Fastq files were aligned to a multi-specie reference genome to generate the SAM/BAM files for each sample. BAM files were filtered and a read count table was prepared. Finally, BAM files were visualized using IGV software and the reads with low percentage of identity wereconfirmed with nBLAST analyses against the NCBI public database. This analysis indicated a wide range of coverage for the highly processed samples tested, from several dozens/hundreds to few millions of reads. The obtained results were compared with the species composition declared on the product label by the manufacturer. Several declared species were identified, as well as no reported species were detected. In addition, nBLAST analyses allowed the addition of new species in the multi-specie reference genome. The present study demonstrates that AgriSeqTM is a viablesolution for genomic applications involving the analysis of hundreds of multiple species in a single sequencing run.